Objective:Critical effective constituents were identified from Bufei Yishen formula(BYF),a traditional herbal compound and combined as effective-constituent compatibility(ECC)of BYF I,which may have potential bioactiv...Objective:Critical effective constituents were identified from Bufei Yishen formula(BYF),a traditional herbal compound and combined as effective-constituent compatibility(ECC)of BYF I,which may have potential bioactive equivalence to BYF.Methods:The active constituents of BYF were identified using four cellular models and categorised into Groups 1(Bufeiqi),2(Bushen),3(Huatan)and 4(Huoxue)according to Chinese medicinal theory.An orthogonal design and a combination method were used to determine the optimal ratios of effective constituents in each group and the ratios of‘‘Groups 1 to 4"according to their pharmacological activity.We also comprehensively assessed bioactive equivalence between the BYF and the ECC of BYF I in a rat model of chronic obstructive pulmonary disease(COPD).Results:We identified 12 active constituents in BYF.The numbers of constituents in Groups 1 to 4 were 3,2,5 and 2,respectively.We identified the optimal ratios of effective constituents within each group.In Group1,total ginsenosides:Astragalus polysaccharide:astragaloside IV ratio was 9:5:2.In Group 2,icariin:schisandrin B ratio was 100:12.5.In Group 3,nobiletin:hesperidin:peimine:peiminine:kaempferol ratio was4:30:6.25:0:0.In Group 4,paeoniflorin:paeonol ratio was 4:1.An orthogonal design was then used to establish the optimal ratios of Group 1,Group 2,Group 3 and Group 4 in ECC of BYF I.The ratio for total ginsenosides:Astragalus polysaccharide:astragaloside IV:icariin:schisandrin B:nobiletin:hesperidin:peimine:paeoniflorin:paeonol was determined to be 22.5:12.5:5:100:12.5:4:30:6.25:25:6.25.A comprehensive evaluation confirmed that ECC of BYF I presented with bioactive equivalence to the original BYF.Conclusion:Based on the ECC of traditional Chinese medicine formula method,the effective constituents of BYF were identified and combined in a fixed ratio as ECC of BYF I that was as effective as BYF itself in treating rats with COPD.展开更多
目的:探讨补肺益肾方治疗慢性阻塞性肺疾病(C O P D)的机制。方法:以A549细胞为研究对象,实验分为NC组(20%对照组大鼠血清)、RAPA组(20%对照组大鼠血清+40nmol/L雷帕霉素)、FS组(20%补肺益肾方大鼠血清)、FS+RAPA组(20%补肺益肾方大鼠血...目的:探讨补肺益肾方治疗慢性阻塞性肺疾病(C O P D)的机制。方法:以A549细胞为研究对象,实验分为NC组(20%对照组大鼠血清)、RAPA组(20%对照组大鼠血清+40nmol/L雷帕霉素)、FS组(20%补肺益肾方大鼠血清)、FS+RAPA组(20%补肺益肾方大鼠血清+40nmol/L雷帕霉素)、CSE组(20%对照组大鼠血清+5%CSE)、CSE+RAPA组(20%对照组大鼠血清+5%CSE+40nmol/L雷帕霉素)、CSE+FS组(5%CSE+20%补肺益肾方大鼠血清)、CSE+FS+RAPA组(5%CSE+20%补肺益肾方大鼠血清+40nmol/L雷帕霉素)。采用qPCR检测PI3K、AKT、mTOR、ULK1、ATG8、ATG10、ATG14 mRNA表达;采用Western blot检测p-PI3K、p-AKT、p-mTOR、p-ULK1、ATG8、ATG10、ATG14蛋白表达。结果:与CSE组比较,CSE+RAPA、CSE+FS及CSE+FS+RAPA组A549细胞PI3K、AKT、mTOR mRNA表达和p-PI3K、p-AKT、p-mTOR蛋白表达显著降低(P<0.01),ATG8、ATG14 mRNA表达和p-ULK1、ATG8、ATG14蛋白表达明显升高(P<0.05,P<0.01)。结论:补肺益肾方治疗COPD的机制可能与调控PI3K/AKT/mTOR信号通路促进细胞自噬有关。展开更多
目的从通路水平探究慢性阻塞性肺疾病有效方药补肺益肾方的干预机制。方法采用LPS诱导巨噬细胞建立炎症反应模型。基于基因集富集分析(Gene set enrichment analysis,GSEA)方法,筛选巨噬细胞炎症反应相关通路,通过富集评分(Normalized e...目的从通路水平探究慢性阻塞性肺疾病有效方药补肺益肾方的干预机制。方法采用LPS诱导巨噬细胞建立炎症反应模型。基于基因集富集分析(Gene set enrichment analysis,GSEA)方法,筛选巨噬细胞炎症反应相关通路,通过富集评分(Normalized enrichment score,NES)筛选药物干预后发生逆转的通路,揭示补肺益肾方及其配伍的干预机制。结果补肺益肾方所含中药的NES为-1377.23,其中补肾配伍的为-485.07、活血配伍的为-351.86、化痰配伍的为-303.71、益气配伍的为-236.59;补肺益肾方显著逆转的通路为213条,其中活血配伍的为184条、补肾配伍的为147条、化痰配伍的为134条、益气配伍的为133条,逆转率分别为75.41%、60.25%、54.92%、54.51%。TGF-βproduction等90条通路在4个配伍中均被显著逆转。Positive regulation of cytokine production involved in inflammatory response等为配伍特异性逆转通路。结论补肺益肾方各配伍组逆转炎症信号通路的强度依次为补肾、活血、化痰、益气配伍,逆转通路数量依次为活血、补肾、化痰、益气。补肺益肾方可通过调控各配伍共性及特异性逆转通路干预炎症反应。展开更多
基金supported by the National Natural Science Fundation of China(No.81973822,81130062,81603473)National Public Welfare Industry Research Project-Traditional Chinese Medicine Industry(No.201507001-01)National Key Research and Development Program of China(No.2018YFC1704806)。
文摘Objective:Critical effective constituents were identified from Bufei Yishen formula(BYF),a traditional herbal compound and combined as effective-constituent compatibility(ECC)of BYF I,which may have potential bioactive equivalence to BYF.Methods:The active constituents of BYF were identified using four cellular models and categorised into Groups 1(Bufeiqi),2(Bushen),3(Huatan)and 4(Huoxue)according to Chinese medicinal theory.An orthogonal design and a combination method were used to determine the optimal ratios of effective constituents in each group and the ratios of‘‘Groups 1 to 4"according to their pharmacological activity.We also comprehensively assessed bioactive equivalence between the BYF and the ECC of BYF I in a rat model of chronic obstructive pulmonary disease(COPD).Results:We identified 12 active constituents in BYF.The numbers of constituents in Groups 1 to 4 were 3,2,5 and 2,respectively.We identified the optimal ratios of effective constituents within each group.In Group1,total ginsenosides:Astragalus polysaccharide:astragaloside IV ratio was 9:5:2.In Group 2,icariin:schisandrin B ratio was 100:12.5.In Group 3,nobiletin:hesperidin:peimine:peiminine:kaempferol ratio was4:30:6.25:0:0.In Group 4,paeoniflorin:paeonol ratio was 4:1.An orthogonal design was then used to establish the optimal ratios of Group 1,Group 2,Group 3 and Group 4 in ECC of BYF I.The ratio for total ginsenosides:Astragalus polysaccharide:astragaloside IV:icariin:schisandrin B:nobiletin:hesperidin:peimine:paeoniflorin:paeonol was determined to be 22.5:12.5:5:100:12.5:4:30:6.25:25:6.25.A comprehensive evaluation confirmed that ECC of BYF I presented with bioactive equivalence to the original BYF.Conclusion:Based on the ECC of traditional Chinese medicine formula method,the effective constituents of BYF were identified and combined in a fixed ratio as ECC of BYF I that was as effective as BYF itself in treating rats with COPD.
文摘目的:探讨补肺益肾方治疗慢性阻塞性肺疾病(C O P D)的机制。方法:以A549细胞为研究对象,实验分为NC组(20%对照组大鼠血清)、RAPA组(20%对照组大鼠血清+40nmol/L雷帕霉素)、FS组(20%补肺益肾方大鼠血清)、FS+RAPA组(20%补肺益肾方大鼠血清+40nmol/L雷帕霉素)、CSE组(20%对照组大鼠血清+5%CSE)、CSE+RAPA组(20%对照组大鼠血清+5%CSE+40nmol/L雷帕霉素)、CSE+FS组(5%CSE+20%补肺益肾方大鼠血清)、CSE+FS+RAPA组(5%CSE+20%补肺益肾方大鼠血清+40nmol/L雷帕霉素)。采用qPCR检测PI3K、AKT、mTOR、ULK1、ATG8、ATG10、ATG14 mRNA表达;采用Western blot检测p-PI3K、p-AKT、p-mTOR、p-ULK1、ATG8、ATG10、ATG14蛋白表达。结果:与CSE组比较,CSE+RAPA、CSE+FS及CSE+FS+RAPA组A549细胞PI3K、AKT、mTOR mRNA表达和p-PI3K、p-AKT、p-mTOR蛋白表达显著降低(P<0.01),ATG8、ATG14 mRNA表达和p-ULK1、ATG8、ATG14蛋白表达明显升高(P<0.05,P<0.01)。结论:补肺益肾方治疗COPD的机制可能与调控PI3K/AKT/mTOR信号通路促进细胞自噬有关。
文摘目的从通路水平探究慢性阻塞性肺疾病有效方药补肺益肾方的干预机制。方法采用LPS诱导巨噬细胞建立炎症反应模型。基于基因集富集分析(Gene set enrichment analysis,GSEA)方法,筛选巨噬细胞炎症反应相关通路,通过富集评分(Normalized enrichment score,NES)筛选药物干预后发生逆转的通路,揭示补肺益肾方及其配伍的干预机制。结果补肺益肾方所含中药的NES为-1377.23,其中补肾配伍的为-485.07、活血配伍的为-351.86、化痰配伍的为-303.71、益气配伍的为-236.59;补肺益肾方显著逆转的通路为213条,其中活血配伍的为184条、补肾配伍的为147条、化痰配伍的为134条、益气配伍的为133条,逆转率分别为75.41%、60.25%、54.92%、54.51%。TGF-βproduction等90条通路在4个配伍中均被显著逆转。Positive regulation of cytokine production involved in inflammatory response等为配伍特异性逆转通路。结论补肺益肾方各配伍组逆转炎症信号通路的强度依次为补肾、活血、化痰、益气配伍,逆转通路数量依次为活血、补肾、化痰、益气。补肺益肾方可通过调控各配伍共性及特异性逆转通路干预炎症反应。
